The CD34+CD38–Isolation Kit, human has been developed to isolate a CD38– subset of CD34+ cells, i.e., primitive hematopoietic stem and progenitor cells, from cord blood, bone marrow, or apheresis.
The CD34+CD38– Cell Isolation Kit has been developed for the isolation of the more primitive CD38– subpopulation of CD34+ cells. CD34 is a well-established marker of human hematopoietic stem and progenitor cells and additionally expressed on hemangioblasts, endothelial progenitor cells, and mature endothelial cells. CD38 is absent on most primitive CD34+ stem cells and upregulated on CD34+ cells upon lymphocyte commitment. CD34+CD38- cells are isolated in a two-step isolation protocol. First, the CD34+ cells are positively enriched using CD34 MultiSort MicroBeads. Subsequently, the MultiSort MicroBeads are enzymatically released from the CD34 antibody. The isolated CD34+ are then labeled with CD38 MicroBeads to deplete CD38+ cells in a second separation step.
- CD34+ CD38– cell expansion in culture
- Lentiviral transduction
- Molecular analysis
- In vitro differentiation of CD34+ CD38– cells
- Generation of humanized mice
CD34+CD38– cells were isolated from human cord blood MNCs by using the CD34 MultiSort MicroBeads, two LS Columns, and a MidiMACS™ Separator for the first separation. In the second separation step, the CD38 MicroBeads, one LS Column, and a MidiMACS Separator have been used. Cells were fluorescently stained with CD34-FITC, CD38-APC, CD45-VioBlue®, Labeling Check Reagent‑APC, and Propidium Iodide Solution and analyzed by flow cytometry using the MACSQuant. Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.
Before CD38 depletion